Clinical efficacy of hematoporphyrin monomethyl ether-based photodynamic therapy for the treatment of port-wine stain and its sonographic changes / 中华皮肤科杂志

Objective@#To evaluate the clinical efficacy of hematoporphyrin monomethyl ether-based photodynamic (HMME-PDT) therapy for the treatment of port-wine stain (PWS) and its sonographic changes.@*Methods@#A total of 45 patients with confirmed PWS were enrolled from the Department of Dermatology, Third Affiliated Hospital of Soochow University from March 2017 to June 2018, including 5 with pink PWS, 39 with purplish red PWS and 1 with thickened PWS. All the patients received 3 sessions of HMME-PDT therapy. The skin thickness and density were compared before and after the treatment by using high-frequency ultrasound. Ranked data were analyzed by using nonparametric test. Measurement data were expressed as mean ± standard deviation, and analyzed using ony-way analysis of variance. Multiple comparisons were performed using Student-Newman-Keuls-q (SNK-q) test. The results were considered to be statistically significant if P < 0.05.@*Results@#Among the 45 patients with PWS who completed the treatment and follow-up, 10 were cured, 21 received marked improvement, 12 received improvement, and 1 showed no response. The total response rate was 97.78%, and the response rate in the patients with pink PWS was higher than that in the patients with purplish red PWS (U = 12.50, P < 0.001) . The difference value of the skin thickness or skin density before and after the treatment significantly differed among the cured patients, patients receiving marked improvement and those receiving improvement (skin thickness:0.65 ± 0.21, 0.56 ± 0.88, 0.37 ± 0.12 mm respectively; skin density: -8.65 ± 2.19, -6.86 ± 2.79, -4.92 ± 2.91 g/cm3 respectively; F = 14.528, 5.428 respectively, both P < 0.001) , and the difference values of the skin thickness and density were significantly higher in the cured patients than in those receiving improvement (q = 5.82, 4.63, both P < 0.05) . Erythematous swelling to different extents occurred at the laser-exposed sites in the zygomatic and cheek region in 23 patients with PWS and in the frontal-zygomatic region in 6 with PWS after the HMME-PDT therapy, but gradually regressed about 1 week later. Pale brown crusts were observed at the laser-exposed sites in 35 patients, and shed spontaneously about 3 weeks later. Post-inflammatory hyperpigmentation at the laser-exposed sites was observed in 4 patients, and gradually regressed after 2-month follow-up.@*Conclusions@#HMME-PDT therapy is effective for the treatment of PWS, with high safety and few adverse reactions. High-frequency ultrasound can be used for objectively evaluating the clinical efficacy of HMME-PDT therapy.

Identification of the targets of hematoporphyrin derivative in lung adenocarcinoma using integrated network analysis

BACKGROUND: Hematoporphyrin derivative (HPD) has a sensibilization effect in lung adenocarcinoma. This study was conducted to identify the target genes of HPD in lung adenocarcinoma. METHODS: RNA sequencing was performed using the lung adenocarcinoma cell line A549 after no treatment or treatment with X-ray or X-ray + HPD. The differentially expressed genes (DEGs) were screened using Mfuzz package by noise-robust soft clustering analysis. Enrichment analysis was carried out using "BioCloud" online tool. Protein-protein interaction (PPI) network and module analyses were performed using Cytoscape software. Using WebGestalt tool and integrated transcription factor platform (ITFP), microRNA target and transcription factor (TF) target pairs were separately predicted. An integrated regulatory network was visualized with Cytoscape software. RESULTS: A total of 815 DEGs in the gene set G1 (continuously dysregulated genes along with changes in processing conditions [untreated-treated with X-ray-X-ray + treated with HPD]) and 464 DEGs in the gene set G2 (significantly dysregulated between X-ray + HPD-treated group and untreated/X-ray-treated group) were screened. The significant module identified from the PPI network for gene set G1 showed that ribosomal protein L3 (RPL3) gene could interact with heat shock protein 90 kDa alpha, class A member 1 (HSP90AA1). TFs AAA domain containing 2 (ATAD2) and protein inhibitor of activated STAT 1 (PIAS1) were separately predicted for the genes in gene set G1 and G2, respectively. In the integrated network for gene set G2, ubiquitin-specific peptidase 25 (USP25) was targeted by miR-200b, miR-200c, and miR-429. CONCLUSION: RPL3, HSP90AA1, ATAD2, and PIAS1 as well as USP25, which is targeted by miR-200b, miR-200c, and miR-429, may be the potential targets of HPD in lung adenocarcinoma.

Clinical efficacy of hematoporphyrin monomethyl ether-based photodynamic therapy for the treatment of port-wine stain and its sonographic changes / 中华皮肤科杂志

Objective To evaluate the clinical efficacy of hematoporphyrin monomethyl etherbased photodynamic (HMME-PDT) therapy for the treatment of port-wine stain (PWS) and its sonographic changes.Methods A total of 45 patients with confirmed PWS were enrolled from the Department of Dermatology,Third Affiliated Hospital of Soochow University from March 2017 to June 2018,including 5 with pink PWS,39 with purplish red PWS and 1 with thickened PWS.All the patients received 3 sessions of HMME-PDT therapy.The skin thickness and density were compared before and after the treatment by using high-frequency ultrasound.Ranked data were analyzed by using nonparametric test.Measurement data were expressed as mean ± standard deviation,and analyzed using ony-way analysis of variance.Multiple comparisons were performed using Student-Newman-Keuls-q (SNK-q) test.The results were considered to be statistically significant if P ＜ 0.05.Results Among the 45 patients with PWS who completed the treatment and follow-up,10 were cured,21 received marked improvement,12 received improvement,and 1 showed no response.The total response rate was 97.78％,and the response rate in the patients with pink PWS was higher than that in the patients with purplish red PWS (U =12.50,P ＜ 0.001).The difference value of the skin thickness or skin density before and after the treatment significantly differed among the cured patients,patients receiving marked improvement and those receiving improvement (skin thickness:0.65 ± 0.21,0.56 ± 0.88,0.37 ± 0.12 mm respectively;skin density:-8.65 ± 2.19,-6.86 ± 2.79,-4.92 ± 2.91 g/cm3 respectively;F =14.528,5.428 respectively,both P ＜ 0.001),and the difference values of the skin thickness and density were significantly higher in the cured patients than in those receiving improvement (q =5.82,4.63,both P ＜ 0.05).Erythematous swelling to different extents occurred at the laser-exposed sites in the zygomatic and cheek region in 23 patients with PWS and in the frontal-zygomatic region in 6 with PWS after the HMME-PDT therapy,but gradually regressed about 1 week later.Pale brown crusts were observed at the laser-exposed sites in 35 patients,and shed spontaneously about 3 weeks later.Post-inflammatory hyperpigmentation at the laser-exposed sites was observed in 4 patients,and gradually regressed after 2-month follow-up.Conclusions HMME-PDT therapy is effective for the treatment of PWS,with high safety and few adverse reactions.High-frequency ultrasound can be used for objectively evaluating the clinical efficacy of HMME-PDT therapy.

Progression of basic research, clinical application of photodynamic therapy and fluorescence-guided surgery in glioma treatment / 中南大学学报(医学版)

The prognosis of glioma remains poor because of the infiltrative nature and the high local relapse rate.The current goals for patients with gliomas is maximal safe resection and adjuvant therapy.Tumor-specific photosensitizer,such as hematoporphyrin derivative (HpD) 5-aminolevulinic acid (5-ALA),can be selective up-taken and accumulated in tumor tissue.Light with appropriate wavelength can penetrate tumor tissue and excite the photosensitizer.The excited photosensitizer within glioma cells permits fluorescence visualization of tumor tissue during surgery and has been introduced in treatment of glioma as fluorescence-guided surgery (FGS).On the other hand,the toxicity of singlet oxygen generated by the excited photosensitizer has been used as photodynamic therapy (PDT) in selective destruction of the tumor.Some reports demonstrated the usefulness of adding PDT as an intraoperative adjuvant therapy,but the complexity associated with its implementation and the introduction of TMZ prevented PDT from becoming a routine therapy.However,FGS using 5-ALA in patients with malignant brain tumors has surfaced globally and may become a useful tool in increasing the extent of resection in gliomas.

Construction and evaluation of charge-reversal nanocarrier loaded with hematoporphyrin monomethyl ether / 药学学报

Charge-reversal nanocarrier was constructed to enhance lysosomal escape and improve an-titumor effect. We synthesized the cholesterol-polyethyleneimine-hexahydrophthalic anhydride (Chol-PEI-HHPA) polymer and characterized by 1H NMR. The charge-reversal liposomes (Lipo-HHPA) were synthesized and the hematoporphyrin monomethyl ether (HMME) was loaded. pH-triggered charge conversion was determined at different pH values. The lysosomal escape and cytotoxicity of the Lipo-HHPA were evaluated in MCF-7 cells. The Lipo-HHPA was uniform with an average particle size of 102 nm. Upon the irradiation of ultrasound, burst release of HMME could be observed. The zeta potential of Lipo-HHPA changed sharply from negative (-23.5 mV) to positive (+21.2 mV) over the pH range of 7.4-4.5. In the cellular uptake experiment, the lysosomal escape of Lipo-HHPA was observed. HMME loaded Lipo-HHPA displayed obviously enhanced cytotoxicity towards MCF-7 cells. These results indicate that the charge-reversal liposomes hold a great potential in improving the cytotoxicity and antitumor effect.

Clinical comparison between photodynamic therapy and pulsed dye laser for the treatment of port wine stains / 中华皮肤科杂志

Objective To compare the clinical efficacy and adverse effects of photodynamic therapy (PDT) versus pulsed dye laser(PDL)for the treatment of port wine stains(PWS). Methods Forty?five patients with PWS were enrolled in this study. The PWS lesions in each patient were randomly divided into PDT and PDL areas. Hematoporphyrin monomethyl ether of 5 mg/kg was injected intravenously into the PDT area protected from light, followed by 20?minute irradiation with a 532?nm, solid?state, continuous?wave laser(power density:80-100 mw/cm2;spot diameter: 7 cm)10 minutes later. The PDL area was treated with a single session of 595?nm pulsed dye laser radiation(spot diameter:7 mm;pulse width:10 ms;energy density:10-12 J/cm2). The interval between PDT and PDL treatment was no shorter than two months. Follow up visits were scheduled on day 4 and week 8 after each treatment. Adverse reactions were recorded, and photographs were taken before and 8 weeks after the treatment for evaluation of lesion regression. Results In the case of PDT area, 10 cases(22.22%)were nearly cured, 22(48.89%)achieved marked improvement, 9(20.00%)improvement, 4(8.89%)no improvement. As far as the PDL area is concerned, 6 cases(13.33%)were nearly cured, 16(35.56%)achieved marked improvement, 18(40.00%)improvement, and 5 (11.11%)no improvement. The response rate was significantly higher in the PDT area than in the PDL area(Z=2.48, P0.05). Conclusion For the treatment of PWS, both PDT and PDL are effective and safe, and single?session PDT appears to be superior to single?session PDL.

Hematoporphyrin monomethyl ether combined with He-Ne laser irradiation-induced apoptosis in canine breast cancer cells through the mitochondrial pathway

Hematoporphyrin monomethyl ether (HMME) combined with He-Ne laser irradiation is a novel and promising photodynamic therapy (PDT)-induced apoptosis that can be applied in vitro on canine breast cancer cells. However, the exact pathway responsible for HMME-PDT in canine breast cancer cells remains unknown. CHMm cells morphology and apoptosis were analyzed using optical microscope, terminal deoxynucleotidyl transferase dUTP nick end labeling fluorescein staining and DNA ladder assays. Apoptotic pathway was further confirmed by Real-time-polymerase chain reaction and Western blotting assays. Our results showed that HMME-PDT induced significant changes in cell morphology, such as formation of cytoplasmic vacuoles and the gradual rounding of cells coupled with decreased size and detachment. DNA fragmentation and cell death was shown to occur in a time-dependent manner. Furthermore, HMME-PDT increased the activities of caspase-9 and caspase-3, and released cytochrome c from mitochondria into the cytoplasm. HMME-PDT also significantly increased both mRNA and protein levels of Bax and decreased P53 gene expression in a time-dependent manner, while the mRNA and protein expression of Bcl-2 were repressed. These alterations suggest that HMME-PDT induced CHMm cell apoptosis via the mitochondrial apoptosis pathway and had anti-canine breast cancer effects in vitro.

Deciphering the binding modes of hematoporphyrin to bovine serum albumin.

Interaction of proteins with small molecules is important in understanding delivery and transport of different therapeutic agents, including drugs. In the present study, we investigated the interaction between hematoporphyrin (HP), the principal component of photosensitizing drug with bovine serum albumin (BSA) in aqueous buffer solution using UV-Vis absorption spectroscopy and fluorescence measurements. The results were further substantiated by molecular docking and molecular dynamics (MD) simulation. Our results revealed that fluorescence of BSA was dominantly quenched by the ground-state complex formation with HP accompanied by the electronic energy transfer (EET) to the later. We experimentally determined the thermodynamic parameters such as G0, H0, and S0 for the HP-BSA system which were -35.5 kJ mole-1, -56.4 kJ mole-1 and -0.06 kJ mole-1 K-1, respectively. These parameters suggested hydrogen-bonding and Van der Waals forces playing major role in the complexation. This was also supported by the binding energy parameters calculated by molecular docking. Moreover, the experimentally determined G0 nicely correlated with those determined by molecular docking and MD-simulation. Further, computational results clearly showed that the binding of HP with BSA in the subdomains IB and IIA.

Inhibitation and radiosensitization effects of hematoporphyrin and radiotherapy on H22 liver cancer mice / 中国生化药物杂志

Objective To observe the inhibitation and radiosensitization effect of hematoporphyrin and radiotherapy on H22 liver cancer mice.Methods Mice models of liver cancer were established by subcutaneous injection of H22 cells,tumor-bearing mice were randomly divided into:control group, simple radiotherapy group, small dose hematoporphyrin with radiotherapy group (combined treatment group Ⅰ), middle dose hematoporphyrin with radiotherapy group(combined treatment group Ⅱ),and high dose hematoporphyrin with radiotherapy group (combined treatment group Ⅲ).Tumor volumes,inhibition rate,sensitizing factor,the survival time of mice,pathology,apoptosis and other indicators in five groups were observed after14 days. Results Hematoporphyrin and radiotherapy can inhibit tumor growth in mice,The q values in three combined treatment groups were all above 1.0 at different time points ,and combined treatment groupⅢwas the highest.The three combined treatment groups had higher apoptosis rate than control group and simple radiotherapy group,but the difference within groups were not statistically significant. The survival time in three combined treatment groups were higher than control group,the combined treatment group Ⅱ was the longest and simple radiotherapy group was the shortest.Conclusion Hematoporphyrin has tumor inhibitition and radiation sensitizing effect on H22 liver cancer mice,and the effect was positive correlation with drug dose. Hematoporphyrin combined with radiotherapy can extend the lifespan of mice in H22 liver cancer,its sensitizing mechanism may be association with induction of H22 cell apoptosis.

Impact of HMME-PDT on Pg of the surface of implants and cementum tablets / 国际生物医学工程杂志

Objective To evaluate the bactericidal effect of diode laser on Porphyromonas gingivalis (Pg),and to explore an optimized protocol for a safe dose of photodynamic therapy (PDT) to eliminate periodontal pathogens as well as the impact on the implant surfaces,so as to provide theoretical and experimental basis for PDT in periimplantitis therapy.Methods Artificial in vitro models were formatted by culturing Pg standard strain and ITI (International Team for Implantology) implants together in CDC broth.Then artificial in vitro models were treated by different doses of hematoporphyrin monomethyl ether (HMME) and different energy density of laser (EDL) for 60 s.The cultures were counted by colony form unit (CFU),and SPSS 17.0 statistical software was used for data statistical analysis to select the best EDL and HMME dose.Finally,ITI implants were observed by scanning electron microscope (SEM) to evaluate the impact of HMME-PDT on Pg of implant surfaces.Results When EDL was 12 J/cm2 and mass concentration of HMME was 25 μg/ml,SEM observations showed that PDT could effectively kill Pg ((13.00±5.00) CFU)without damaging the implant surfaces.Conclusions PDT therapy combining 630 nm diode laser with photosensitizer HMME have good bactericidal effect on Pg,and the EDL and HMME dose is as small as the clinical applicable safe dose.

Dose effect of laser irradiation in photodynamic therapy for laryngocarcinoma in BALB/c mouse / 国际生物医学工程杂志

Objective The dose and type of light and photosensitizer could seriously affect the curative effect of photodynamic therapy (PDT).The purpose of this study was to observe whether or not PDT with hematoporphyrin monomethyl ether (HMME) can cure laryngocarcinoma in the solid tumor model,and to define the proper laser amount for killing the cancer cells.Methods Forty eight BALB/c mouse models with subcutaneous Hep-2 laryngeal carcinomas were prepared.Mice were divided into six groups depending on the amount of laser received from 30 J/cm2 to 480 J/cm2 including a control group,tumor size in each group was between 8 mm and 10 mm.Tail vein injection were given with HMME prior to applying the laser light,and then illumination was carried out on the tumor at 3 h after HMME administration.Tumor volume,animal weight and histopathologic changes were observed after PDT.Results All mice apparently showed positive results via PDT,and the cancer had been cured in 120 J/cm2 and 480 J/cm2 groups.The laryngeal cancer lesions began to form scab 1 d after PDT and the scab became hard and black after 5 d.The tumor regression began simultaneously and completed around 30 d after PDT.Conclusions PDT may treat laryngeal cancers which sized less than 10 mm in mouse models.The optimum energy to destruct the laryngeal cancer cells may be 120 J/cm2.

Effect of photodynamic therapy on the phosphorylation of Smad3 in hyperplastic scar fibroblasts / 中华皮肤科杂志

Objective To observe the phosphorylation of Smad3 in hyperplastic scar fibroblasts (HSFs) induced by hematoporphyrin monomerthyl ether (HMME) followed by photodynamic therapy (PDT).Methods Fibroblasts were isolated from the hypertrophic scar tissues of 10 patients and subjected to culture in vitro.After 3-5 passages,the HSFs were divided into 4 groups:control group receiving no treatment,PDT group pretreated with HMME of 4 μg/ml followed by PDT,HMME group induced by HMME alone,and laser group irradiated with laser alone.Fluorescence microscopy was used to observe the expression of Smad3 after immunofluorescent staining with anti-Smad3 antibody,and Western blot to detect the expression of Smad3 and phosphorylated Smad3 in these HSFs.Paired t test was conducted to compare the difference in Smad3 and phosphorylated Smad3 expression between these groups.Results The total fluorescence intensity of Smad3 was similar between these groups,but the intranuclear fluorescence signal was significantly weaker in the PDT group than in the control group.The level of phosphorylated Smad3 was statistically decreased in the PDT group compared with the control group (0.20 ± 0.02 vs.0.92 ± 0.15,P ＜ 0.05),but no significant difference was observed between the HMME group and laser group (P ＞ 0.05).Conclusion PDT may inhibit the proliferation of HSFs via attenuating the phosphorylation of Smad3.

Antibacterial efficacy of HMME-PDT on Enterococcus faecalis within infected simulated lateral canals in vitro model with different energy / 国际生物医学工程杂志

Objective The purpose of this study was to assess the antibacterial effects of hematoporphyrin monomethylether-photodynamic therapy (HMME-PDT) on Enterococcus faecalis within infected simulated lateral canals in vitro with different energy.Methods Simulated lateral canals were prepared on extracted teeth.The specimens were infected with Enterococcus faecalis and then were randomly divided into eight groups.Group A was considered as negative control,its specimens were irrigated with physiological saline.Group B was the positive control,they were irrigated with 5.25％ NaClO.Other groups were incubated with HMME at concentration of 40 μg/ml for 5 min,followed by exposure to light at 532 nm for 120 s with different powers in a spiral pattern.Groups were named C-H,in corresponding with the power 50,60,70,80,90,100 mW.Microbial samples (the dentin chips from simulated lateral canals) were taken before and after the treatments.The survival fractions in each simulated lateral canal was calculated by counting colony-forming units(CFUs).Results 1.The HMME-PDT-treated groups resulted in a significant reduction in the number of E.faecalis in simulated lateral canals compared with the negative controls(P＜0.05).2.The antibacterial effects of group D、E、F、G、H were higher than the NaCl0-irrigated group(P＜0.05).3.The difference of antibacterial effects between the F、G、H groups was in significant(P＞0.05).Conclusion HMME-PDT had significant inhibitive efficacy on Enterococcus faecalis within infected simulated lateral canals and the efficacy was power-depended.It can play the best effect with the power of 80 mW.

Comparison of absorption characteristics of hematoporphyrin monomethyl ether (HMME) by ECV304 cells versus HaCaT cells / 中华皮肤科杂志

Objective To compare the absorption characteristics of HMME by a human umbilical vein endothelial cell line ECV 304 versus a human keratinocyte cell line HaCaT.Methods Exponentially growing ECV304 and HaCaT cells were incubated with various concentrations (50,100,150,200 and 250 mg/L) of HMME for 16 h or HMME of 150 mg/L for various durations (15 min,30 min,1 h,3 h,8 h,12 h and 24 h).The quantity of HMME absorbed by the cells were determined by laser scanning confocal microscopy (LSCM).Results The fluorescence intensity was 74.00,125.57,135.24,141.99 and 132.09 for ECV304 cells,93.88,102.45,112.59,108.23 and 104.70 for HaCaT cells,after incubation with HMME of 50,100,150,200 and 250 mg/L,respectively.After treated with HMME of 150 mg/L for 15 min,30 min,1 h,3 h,8 h,12 h and 24 h,ECV304 cells showed a fluorescence intensity of 95.07,103.97,105.96,108.99,112.93,115.36 and 122.91,respectively,and HaCaT cells displayed a fluorescence intensity of 104.25,106.60,108.72,113.75,117.66,114.90 and 118.14,respectively.Conculsions Within a defined range of concentration and duration,the absorption of HMME by both ECV304 and HaCaT cells is,to some extent,concentration- and time-dependent.

Investigating the killing effect of photodynamic therapy induced by different photosensitizers on human leukemia cell in vitro / 国际生物医学工程杂志

Objective To investigate and compare the killing effect of photodynamic therapy (PDT)induced by hematoporphyrin derivative (HpD),hematoporphyrin monomethyl ether (HMME) and photocarcinorin (PsD007) on human leukemia cells K562 in vitro.Methods Human leukemia cells were cultured with serial concentrations of photosensitizers followed by irradiation of different dosage of laser light,then MTT colorimetric assay was applied to measure the relative survival rate of PDT for the cells.Results Significant difference in the inhibitory between the PDT group and control group was observed (P＜0.05).The survival rate of PDT for the cells elevated along with the increase in the concentration of sensitizer and dose of laser light.When the photosensitizer concentration was bigger (25 μg/ml) or the energy density was bigger (7.2 J/cm2),the effect of PsD007 was better than HMME,and they were significantly better than HpD (P＜0.05).Conclusion PDT has significant killing effect on human leukemia cells K562,and its relative inhibitory rate appears to be correlated with the dose of sensitizer and laser light irritation.The effect of PDT is related to the photosensitizers.The effect of HpD-PDT is not as effective as PsD007 and HMME.On the conditions of higher energy density and larger photosensitizer concentration,the effect of PsD007-PDT is better than HMME-PDT.

Detection of IL-2 and IL-6 in human breast tumor Bcap-37 cells treated by hematoporphyrin nonomethyl ether mediated photodynamic therapy / 肿瘤研究与临床

Objective To observe IL-2 and IL-6 changes in the breast tumor Bcap-37 cells reated by hematoporphyrin nonomethyl ether mediated photodynamic therapy (HMME-PDT).Methods Cells in logarithmic growth phase were collected among breast cancer cells cultured in conventional methods.According to blank control group or the experimental group (laser irradiation group,photosensitive agent group and HMME-PDT group),PDT in addition to HMME and HMME-PDT were conducted.The changes of IL-2,IL-6 were detected by radioimmunoassay.Results After HMME-PDT,IL-2 was increased as time passed.After 12,24 and 48 h,compared with IL-2 level in the control group,in laser irradiation group or photosensitive agent group,the levels of IL-2 in HMME-PDT group was significantly differences (P ＜ 0.05).But IL-6 levels decreased.The most obvious changes of IL-6 levels happened at 12h and 24h.There was significant differences between IL-6 in HMME-PDT group with the control group,laser irradiation group or photosensitive agent group (P ＜ 0.05).Conclusion HMME-PDT maybe have destruction effect by altering IL-2,IL-6 activity on breast tumor cells,which provides objective indicators for clinical patients to regulate immune function and auxiliary diagnosis.

The inhibitory effect of photodynamic therapy and of an anti-VCAM-1 monoclonal antibody on the in vivo growth of C6 glioma xenografts

We investigated the effect of photodynamic therapy (PDT) and of an anti-vascular cell adhesion molecule-1 (VCAM-1) monoclonal antibody on the in vivo growth of C6 glioma. Seven days after inoculation with C6 cells, adult male Wistar rats weighing 280-300 g with MRI-confirmed glioma were randomly assigned to 4 groups (N = 15 per group): PDT + VCAM-1 antibody group; PDT group; VCAM-1 antibody group; control group. Eight days after inoculation, hematoporphyrin monomethyl ether (HMME) was administered as a photosensitizer and PDT was performed at 630 nm (illumination intensity: 360 J/cm²) for 10 min. VCAM-1 antibody (50 µg/mL) was then administered (0.5 mL) through the tail vein every other day from day 8 to day 16. At day 21, 5 rats in each group were sacrificed and cancers were harvested for immunohistochemistry and Western blot assay for the detection of VCAM-1, and TUNEL assay was used to detect apoptosis. Survival and tumor volume were recorded in the remaining 10 rats in each group. In the PDT group, tumor growth was significantly suppressed (67.2 percent) and survival prolonged (89.3 percent), accompanied by an increase in apoptosis (369.5 percent), when compared to control. Furthermore, these changes were more pronounced in the PDT + VCAM-1 antibody group. After PDT, VCAM-1 expression was markedly increased (121.8 percent) and after VCAM-1 monoclonal antibody treatment, VCAM-1 expression was significantly reduced (58.2 percent). PDT in combination with VCAM-1 antibody can significantly inhibit the growth of C6 glioma and prolong survival. This approach may represent a promising strategy in the treatment of glioma.

Effects of caspase 3 on hypertrophic scar fibroblasts in hematoporphyrin monomerthyl etherphotodynamic therapy (HMME-PDT) / 中华皮肤科杂志

ObjectiveTo investigate the role of caspase 3 in HMME-induced apoptosis in hypertrophic scar fibroblasts (HSFs).MethodsFibroblasts were obtained from 10 patients with untreated hypertrophic scar,and subjected to a primary culture.After 4 to 6 passages of culture,the HSFs were divided into 3 groups to remain untreated(control group),be treated with HMME followed by photodynamic therapy (HMME-PDT group),or the combination of HMME and Z-DEVD-FMK followed by photodynamic therapy (caspase 3 inhibitor group).At 12 hours after the therapy,HSFs were collected and immunofluorescence microscopy was used to observe the fluorescence intensity of caspase 3 after staining with fluorescein isocyanate (FITC) and popodium iodide (PI),flow cytometry was performed to determine the percentage of caspase 3-positive HSFs and apoptosis rate in HSFs after single staining with FITC and PI respectively.Results The fluorescence intensity of caspase 3 was weak in the control group and caspase 3 inhibitor group,but was strong in the HMME-PDT group.An increased percentage of caspase 3-positive HSFs was noted in the HMMEPDT group compared with the control group and caspase 3 inhibitor group(30.86％ ± 1.21％ vs.3.12％ ±0.28％ and 2.46％ ± 0.18％,t =19.92,21.76,both P ＜ 0.05).The apoptosis rate in HSFs was significantly higher in the HMME-PDT group and caspase 3 inhibitor group than in the control group(30.54％ ± 3.78％ and 10.46％ ± 2.15％ vs.2.45％ ± 0.22％,t =35.90,27.97,both P＜ 0.05),and higher in the HMME-PDT group than in the caspase 3 inhibitor group.ConclusionsThe apoptosis in HSFs induced by HMME-PDT is closely related to the activation of caspase 3,while caspase 3 seems to be dispensable for the apoptosis.

Apoptosis of hypertrophic scar fibroblasts induced by HMME-PDT / 中华医学美学美容杂志

Objective To investigate the apoptotic effects of hypertrophic scar fibroblast (HSF) induced by HMME-PDT.Methods Fibroblasts were cultured from nontreated hypertrophic scars,and cells at passages 4-6 were used for the experiments (photosensitizer dose 4 μg/ml,λ630 nm,pow er density 10 mw/cm2,energy fluence 2.5 J/cm2).Morphological and biochemical changes in fibroblasts were assessed by Hoechst 33258 staining and fluorescence microscopy.The rate of apoptotic or necrotic cells was detected by flow cytometry (FCM) through double staining of Annexin V -FITC and popodium iodide (PI),respectively.Results Marked morphological features of cell apoptosis were viewed under the fluorescent microscope through Hoechst 33258 staining.The analysis of FCM indica ted that the apoptotic rate was significantly increased after HMME PDT [(34.82 ± I.42) ％ vs (3.12±0.28) ％,P＜0.05],and apoptotic rate was higher than necrosis rate [(14.65±1.02) ％ vs (34.82±1.42) ％,P＜0.05].Conclusions Low level exposure to 630 nm PDT mediated by HMME appears to induce fibroblast apoptosis.

Photodynamic inactivation of four Candida species induced by photogem®

This study evaluated the in vitro susceptibility of C. albicans, C. dubliniensis, C. tropicalis and C. krusei to photodynamic therapy (PDT) induced by Photogem® and light emitting diode (LED). Suspensions of each Candida strain were treated with three photosensitizer (PS) concentrations (10, 25 and 50 mg/L) and exposed to 18.0, 25.5 and 37.5 J/cm² LED light fluences (λ ~ 455 nm). Control suspensions were treated only with PS concentrations, only exposed to the LED light fluences or not exposed to LED light or PS. Sixteen experimental conditions were obtained and each condition was repeated three times. From each sample, serial dilutions were obtained and aliquots were plated on Sabouraud Dextrose Agar. After incubation of plates (37 ºC for 48 hours), colonies were counted (cfu/mL) and the data were statistically analyzed by ANOVA and the Tukey test (α=0.05). Complete killing of C. albicans was observed after 18.0 J/cm² in association with 50 mg/L of PS. C. dubliniensis were inactivated after 18.0 J/cm² using 25 mg/L of PS. The inactivation of C. tropicalis was observed after photosensitization with 25 mg/L and subsequent illumination at 25.5 J/cm². For C. krusei, none of the associations between PS and light resulted in complete killing of this species. PDT proved to be effective for the inactivation of C. albicans, C. dubliniensis and C. tropicalis. In addition, reduction in the viability of C. krusei was achieved with some of the PS and light associations.